常温机械灌注修复长时间冷保存供肝的大动物实验研究

目的探索常温机械灌注(NMP)在挽救大动物边缘供肝中的价值。 方法6只雄性、10～12月龄巴马小型猪分为静态冷保存(SCS)6 h组和SCS 24 h组，每组3只，分别于供肝获取后SCS 6 h、24 h后进行2 h NMP复苏。基于荷兰Organ Assist公司Liver Assist系统和供体猪自身血液，整合四通道生理仪器及自制灌注管路搭建NMP平台。在NMP过程中分别于灌注0、15、60、90、120 min 5个时间点收集灌注液用于肝功能(ALT、AST)检测及血气分析(pH、氧分压)。灌注结束取肝左叶相同位置少许组织，以10%甲醛固定，用于后续HE染色。 结果NMP开始时，SCS 24 h组ALT、AST水平略高于SCS 6 h组，之后AST、ALT水平均缓慢上升。NMP开始时SCS 24 h组pH值中位数为7.28，氧分压中位数为46 mmHg(1 mmHg＝0.133 kPa，下同)，均低于SCS 6 h组(7.36，52 mmHg)，经过2 h NMP后SCS 24 h组pH值和氧分压逐渐接近甚至优于SCS 6 h组。HE染色发现，灌注前SCS 24 h组肝脏充血严重，炎性细胞浸润明显；NMP 2 h后，肝脏充血明显改善、炎性细胞减少，与SCS 6 h组无明显差异。 结论基于Liver Assist系统和供体猪自身血液成功搭建的NMP平台，可有效改善长时间冷保存的边缘供肝质量，提示NMP在修复边缘供肝、扩展供肝来源方面具有重要临床及基础研究价值。

Normothermic machine perfusion improves long time cold storage liver quality in a porcine model

ObjectiveTo explore the application value of normothermic machine perfusion (NMP) for marginal liver in a porcine model. MethodsSix healthy male Ba-Ma mini pigs (10-12 months old) were equally divided into two groups according to the experimental design: static cold storage 6 hours (SCS 6 h) followed by 2 h NMP (SCS 6 h+ NMP 2 h) group and (SCS 24 h+ NMP 2 h) group. The NMP perfusion platform was set up based on Liver Assist System, blood of donor, four-channel physiological instruments and self-made perfusion cannula integration. The perfusate of the two groups were collected at 0, 15, 60, 90 and 120 min during NMP, which were used for liver function test and arterial blood gas analysis. A little liver tissue (fixed by 10% formaldehyde) was obtained from the left lobe, which was used for HE staining. ResultsThe level of ALT and AST were higher in (SCS 24 h+ NMP 2 h) group than those of (SCS 6 h+ NMP 2 h) group at the beginning of NMP, and both rise gradually in the two groups after perfusion. The median pH and oxygen partial pressure value of (SCS 24 h+ NMP 2 h) group were 7.28 and 46 mmHg at the beginning of NMP, both higher than those in (SCS 6 h+ NMP 2 h) group (7.36, 52 mmHg), but the above indexes in (SCS 24 h+ NMP 2 h) group are progressively closer to or even better than those of (SCS 6 h+ NMP 2 h) group. Pathology results showed that liver congestion and inflammatory cell infiltration were serious in (SCS 24 h+ NMP 2 h) group before NMP, whereas after perfusion, these symptoms had improved markedly without significant difference to (SCS 6 h+ NMP 2h) group. ConclusionThe NMP perfusion platform based on Liver Assist System and blood of donor could effective improve the quality of long-time cold preservation marginal livers, which means that it had important clinical and fundamental research value for marginal livers repairment and donor liver pool enlargement.